mitochondrial ribosomal protein L4 Overexpression Lysate Summary
                         
                                
                                
                                
            | Description | 
            mitochondrial ribosomal protein L4 Transient Overexpression Lysate Expression Host: HEK293T
  Plasmid: RC200861
  Accession#: NM_146388
  Protein Tag: C-MYC/DDK
   You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. | 
        
            | Gene | 
            MRPL4  | 
        
                                
                          Applications/Dilutions
                                
                                    
                                    
                                        
                              
                                  | Dilutions | 
                                  
                                      
                                   | 
                              
            | Application Notes | 
            This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.  | 
        
            | Theoretical MW | 
            29.3 kDa.  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.  | 
        
                                    
                                  Packaging, Storage & Formulations
            | Storage | 
            Store at -80C. Avoid freeze-thaw cycles.  | 
        
            | Buffer | 
            RIPA buffer  | 
        
Lysate Details for Array
                 Notes
                    
                        HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
                     Alternate Names for mitochondrial ribosomal protein L4 Overexpression Lysate
                     Background
 
                    
                    Mammalian mitochondrial ribosomal proteins are encoded by nuclear genes and help in protein synthesis within the mitochondrion. Mitochondrial ribosomes (mitoribosomes) consist of a small 28S subunit and a large 39S subunit. They have an estimated 75% protein to rRNA composition compared to prokaryotic ribosomes, where this ratio is reversed. Another difference between mammalian mitoribosomes and prokaryotic ribosomes is that the latter contain a 5S rRNA. Among different species, the proteins comprising the mitoribosome differ greatly in sequence, and sometimes in biochemical properties, which prevents easy recognition by sequence homology. This gene encodes a 39S subunit protein. Sequence analysis identified alternatively spliced variants that encode different protein isoforms.  Transcript Variant: This variant (3) uses an alternate splice site in the 3' coding region, compared to variant 1, that results in a frameshift. It encodes an isoform (b) which has a shorter and distinct C-terminus compared to isoform a.  Sequence Note: The RefSeq transcript and protein were derived from genomic sequence to make the sequence consistent with the reference genome assembly. The genomic coordinates used for the transcript record were based on alignments.
                      Limitations
 
                    
                    This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are 
guaranteed for 6 months from date of receipt.
 
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